Ameliorative Effect of Ethanolic Extract of Moringa oleifera Leaves in Combination with Curcumin against PTZ-Induced Kindled Epilepsy in Rats: In Vivo and In Silico

Abstract

The ameliorative effect of ethanolic extract of M. oleifera (MOEE) leaves in combination with curcumin against seizures, cognitive impairment, and oxidative stress in the molecular docking of PTZ-induced kindled rats was performed to predict the potential phytochemical effects of MOEE and curcumin against epilepsy. The effect of pretreatment with leaves of M. oleifera ethanolic extracts (MOEE) (250 mg/kg and 500 mg/kg, orally), curcumin (200 mg/kg and 300 mg/kg, orally), valproic acid used as a standard (100 mg/kg), and the combined effect of MOEE (250 mg/kg) and curcumin (200 mg/kg) at a low dose on Pentylenetetrazole was used for (PTZ)-induced kindling For the development of kindling, individual Wistar rats (male) were injected with pentyletetrazole (40 mg/kg, i.p.) on every alternate day. Molecular docking was performed by the Auto Dock 4.2 tool to merge the ligand orientations in the binding cavity. From the RCSB website, the crystal structure of human glutathione reductase (PDB ID: 3DK9) was obtained. Curcumin and M. oleifera ethanolic extracts (MOEE) showed dose-dependent effects. The combined effects of MOEE and curcumin leaves significantly improved the seizure score and decreased the number of myoclonic jerks compared with a standard dose of valproic acid. PTZ kindling induced significant oxidative stress and cognitive impairment, which was reversed by pretreatment with MOEE and curcumin. Glutathione reductase (GR) is an enzyme that plays a key role in the cellular control of reactive oxygen species (ROS). Therefore, activating GR can uplift antioxidant properties, which leads to the inhibition of ROS-induced cell death in the brain. The combination of the ethanolic extract of M. oleifera (MOEE) leaves and curcumin has shown better results than any other combination for antiepileptic effects by virtue of antioxidant effects. As per the docking study, chlorogenic acid and quercetin treated with acombination of curcumin have much more potential.

Keywords: Moringa oleifera; Pentylenetetrazole; curcumin; neuroprotective; oxidative stress.

Figure 1

Effect of M. oleifera ethanolic extract (MOEE) and curcumin on the seizure severity score in PTZ-treated Wistar albino rats. Each value is expressed as the mean ± SEM; number of animal = 6.

Figure 2

Effect of MOEE and curcumin on elevated plus maze apparatus in PTZ-induced kindled rats. Values areexpressed as the mean ± standard error of the mean (SEM).* p < 0.01, ** p < 0.001, *** p < 0.0001, a—control vs. PTZ, b—PTZ vs. all groups.

Figure 3

Effect of MOEE and curcumin on the step through latency test in PTZ-induced kindled Rats. Valuesareexpressed as the mean ± standard error of the mean. ** p < 0.001, *** p < 0.0001, a—control vs. PTZ, b—PTZ vs. all groups.

Figure 4

Effect of MOEE and curcumin on open field test in PTZ-induced kindled rats. ns = non-significant, a—control vs. PTZ, b—PTZ vs. all groups.

Figure 5

Effect of MOEE and curcumin on thebrain for (A) MDA levels, (B) Glutathione levels, (C) SOD levels, (D) Catalase levels, (E) Nitric oxide levels, and (F) AChE levels in PTZ-induced kindled Rats. Value was expressed as mean ± Standard error of mean (SEM).* p < 0.01, ** p < 0.001, *** p < 0.0001. ns = not significant, a—control vs. PTZ, b—PTZ vs. all groups.

Figure 6

The overlapping of the whole ligand [chlorogenic acid (Saffron color); quercetin (black); curcumin (olive green); internal ligand (pink); valproic acid (Yellow)] in the active site of human glutathione reductase (PDB ID: 3DK9).

Figure 7

(a).Binding interaction of chlorogenic acid (saffron color) in the active site of human glutathione reductase (PDB ID: 3DK9); (b)—Binding interaction of quercetin (black); (c)—Curcumin (olive green); (d)—Valproic Acid (yellow) in the active site of human glutathione reductase.

Figure 7

(a).Binding interaction of chlorogenic acid (saffron color) in the active site of human glutathione reductase (PDB ID: 3DK9); (b)—Binding interaction of quercetin (black); (c)—Curcumin (olive green); (d)—Valproic Acid (yellow) in the active site of human glutathione reductase.

Figure 8

Experimental protocol of Pentylenetetrazole (PTZ)-induced kindling.